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Chapter 8 : Function and Expression of Surface Factors

Authors: Klaus Heuner1, Michael Steinert1, Claudia Dietrich1, Rolf Köhler1, Jörg Hacker1, Gunter Fischer2
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Affiliations: 1: Institut fur Molekulare Infektionsbiologie, Julius-Maximilians Universitat Wiirzburg, Röntgenring 11, D-97070 Wiirzburg, Germany; 2: Forschungsstelle “Enzymologie der Proteinfaltung” der Max-Planck-Gesellschaft, Weinbergweg 22, D-06120 Halle, Germany
Content Type: Monograph
Publication Year: 2002

Category: Bacterial Pathogenesis

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Abstract:

This chapter focuses on the major infectivity potentiator protein (Mip) protein and the function and expression of the flagellum of . The Mip protein forms a homodimer on the bacterial surface; each monomer consists of a proximal and a peripheral domain. The contact regions between the two monomers seem to be located at the N-terminal part of the protein. Motility is an important factor to find a new host for another cycle of intracellular replication and for colonization of new habitats. Flagellated bacteria have been found in lung alveolar spaces of patients with legionellosis. It was shown that expression of the virulent phenotype and motility is regulated coordinately. A gentamicin infection assay revealed a clear difference in the number of intracellular bacteria at the onset of multiplication. is found in very different habitats and it is able to replicate intracellularly in many host cells. Researchers recently demonstrated that regulation of expression is modulated by different environmental factors, such as temperature, growth phase, osmolarity, viscosity, and the nutrient stage.

Citation: Heuner K, Steinert M, Dietrich C, Köhler R, Hacker J, Fischer G. 2002. Function and Expression of Surface Factors, p 43-48. In Marre R, Abu Kwaik Y, Bartlett C, Cianciotto N, Fields B, Frosch M, Hacker J, Lück P (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555817985.ch8
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Function and Expression of Legionella pneumophila Surface Factors
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Citation: Heuner K, Steinert M, Dietrich C, Köhler R, Hacker J, Fischer G. 2002. Function and Expression of Surface Factors, p 43-48. In Marre R, Abu Kwaik Y, Bartlett C, Cianciotto N, Fields B, Frosch M, Hacker J, Lück P (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555817985.ch8
/content/10.1128/9781555817985.chap8.fig8-1
FIGURE 1

Electron micrographs showing the flagellated wild-type strain Corby (A) and the nonflagellated mutant strain KH3 (B). Bacteria were grown to stationary phase at 30°C, suspended in water, and applied to Formvar coated copper grids. Samples were shadowed with platinum-palladium and examined with a Zeiss 10A transmission electron microscope. Bars, 0.5 μm.

10.1128/9781555817985/fig8-1_thmb.gif
10.1128/9781555817985/fig8-1.gif
Image of FIGURE 1
FIGURE 1

Electron micrographs showing the flagellated wild-type strain Corby (A) and the nonflagellated mutant strain KH3 (B). Bacteria were grown to stationary phase at 30°C, suspended in water, and applied to Formvar coated copper grids. Samples were shadowed with platinum-palladium and examined with a Zeiss 10A transmission electron microscope. Bars, 0.5 μm.

Citation: Heuner K, Steinert M, Dietrich C, Köhler R, Hacker J, Fischer G. 2002. Function and Expression of Surface Factors, p 43-48. In Marre R, Abu Kwaik Y, Bartlett C, Cianciotto N, Fields B, Frosch M, Hacker J, Lück P (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555817985.ch8
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FIGURE 2

Gentamicin assay of Corby wild-type, the mutant strain (KH3), and the complemented mutant strain (CD 10) with (A) and HL-60 cells (B). Host cells were incubated with legionellae at a multiplicity of infection of 10 for 2 h. Extracellular bacteria were killed by incubation with gentamicin (80 g/ml) for 1 h, and the CFUs were determined by plating on ABCYE agar plates ( = 0). The rate of intracellular multiplication ( = 0 to = 24) is given above or beneath the error bars. Error bars indicate the standard deviation obtained from three independent experiments.

10.1128/9781555817985/fig8-2_thmb.gif
10.1128/9781555817985/fig8-2.gif
Image of FIGURE 2
FIGURE 2

Gentamicin assay of Corby wild-type, the mutant strain (KH3), and the complemented mutant strain (CD 10) with (A) and HL-60 cells (B). Host cells were incubated with legionellae at a multiplicity of infection of 10 for 2 h. Extracellular bacteria were killed by incubation with gentamicin (80 g/ml) for 1 h, and the CFUs were determined by plating on ABCYE agar plates ( = 0). The rate of intracellular multiplication ( = 0 to = 24) is given above or beneath the error bars. Error bars indicate the standard deviation obtained from three independent experiments.

Citation: Heuner K, Steinert M, Dietrich C, Köhler R, Hacker J, Fischer G. 2002. Function and Expression of Surface Factors, p 43-48. In Marre R, Abu Kwaik Y, Bartlett C, Cianciotto N, Fields B, Frosch M, Hacker J, Lück P (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555817985.ch8
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FIGURE 3

Effects of different environmental factors on the flagellin expression of Corby (pKH23, fusion). was grown in YEB medium at 37°C supplemented with 1% glucose, 30 mM serine, 200 mM sucrose (osmolarity), or polyvinylpyrrolidone (PVP) (viscosity). Cells were harvested at the late exponential growth phase and luciferase activity was measured and is given in relative light units (RLU). Error bars indicate the standard deviation obtained from three independent experiments. □, control; , 1% glucose; , 30 mM serine; , 200 mM sucrose; , 6% PVP.

10.1128/9781555817985/fig8-3_thmb.gif
10.1128/9781555817985/fig8-3.gif
Image of FIGURE 3
FIGURE 3

Effects of different environmental factors on the flagellin expression of Corby (pKH23, fusion). was grown in YEB medium at 37°C supplemented with 1% glucose, 30 mM serine, 200 mM sucrose (osmolarity), or polyvinylpyrrolidone (PVP) (viscosity). Cells were harvested at the late exponential growth phase and luciferase activity was measured and is given in relative light units (RLU). Error bars indicate the standard deviation obtained from three independent experiments. □, control; , 1% glucose; , 30 mM serine; , 200 mM sucrose; , 6% PVP.

Citation: Heuner K, Steinert M, Dietrich C, Köhler R, Hacker J, Fischer G. 2002. Function and Expression of Surface Factors, p 43-48. In Marre R, Abu Kwaik Y, Bartlett C, Cianciotto N, Fields B, Frosch M, Hacker J, Lück P (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555817985.ch8
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References

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1. Bellinger-Kawahara, C.,, and M. A. Horwitz. 1990. Complement component C3 fixes selectively to the major outer membrane protein (MOMP) of Legionella pneumophila and mediates phagocytosis of liposome-MOMP complexes by human monocytes. J. Exp. Med. 172: 1201– 1210.
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